KR102545412B1 - Vaccine Composition for SFTS virus - Google Patents

Abstract

The present invention is a recombinant protein in which the extracellular domain of the Gn or Gc protein of the SFTS virus is fused to the Fc region of a human antibody, NP protein (protein of full-length sequence), or a mixture thereof in type 1 interferon receptor KO mice. A vaccine composition against SFTS virus using Gn or Gc protein or NP protein or a mixture thereof based on the experimental results providing protective immunity is disclosed.

Description

Vaccine composition for severe fever with thrombocytopenia syndrome virus {Vaccine Composition for SFTS virus}

The present invention relates to a vaccine composition for a vaccine composition against severe fever with thrombocytopenia syndrome virus.

Severe fever with thrombocytopenia syndrome (SFTS) is an infectious disease caused by infection with the SFTS virus (SFTSV). Since it was first reported in China in 2009, outbreaks have been reported in Asian countries such as China, Korea, Japan, and Vietnam. SFTSV has three segmented RNAs in its genome, each encoding an RNA polymerase (RdRp), two outer membrane glycoproteins (Gn, Gc), and a nucleocapsid (NP) and non-structural protein (NS). Infection with this virus is accompanied by high fever, gastrointestinal complications, leukopenia, and thrombocytopenia, and the mortality rate is known to be 5 to 20% ( Lancet Infect Dis 2018, 18 , 1127-1137, doi:10.1016/S1473-3099(18 )30293-7). There is no cure yet, and vaccine development is ongoing.

SFTS vaccine development is in the early research stage, and recent reports have confirmed that vaccines using recombinant viral vectors or DNA vaccines can provide protective immunity in infected animal models ( NPJ Vaccines 2019, 4, 5, doi :10.1038/s41541-018-0096-y; Nat Commun 2019, 10, 3836, doi:10.1038/s41467-019-11815-4). It was confirmed that the vaccine using the NS protein induced antibody production, but failed to provide protective immunity in a mouse infection model ( Viral Immunol 2015, 28, 113-122, doi: 10.1089/vim.2014.0100). It has also been reported that immunization with DNA encoding NP and NS genes using a microneedle induces a T cell response specific to these antigens, but it has not been clarified whether protective immunity is provided in animal infection models ( Colloids Surf B Biointerfaces 2017, 159, 54-61, doi:10.1016/j.colsurfb.2017.07.059). Recently, once immunized with live attenuated recombinant vesicular stomatitis virus (rVSV) expressing Gn and Gc proteins, neutralizing antibody production can be induced, and SFTSV-infected type 1 interferon receptor knock-out A paper has been reported that complete protective immunity is induced in (KO) mice ( NPJ Vac cines 2019, 4, 5, doi:10.1038/s41541-018-0096-y). In addition, when immunized with DNA expressing each of the 5 genes of SFTSV, when immunized with two DNAs encoding Gn and Gc, and when immunized with three DNAs encoding NP, NS, and RdRp, SFTSV It has also been reported that protective immunity against lethal infection is conferred in susceptible older weasels (more than 4 years) ( Nat Commun 2019, 10, 3836, doi:10.1038/s41467-019-11815-4). However, it has not yet been reported how much protective immunity is provided when each structural protein (Gn, Gc, NP) is individually immunized, and studies on the efficacy of subunit protein vaccines using these proteins have been published. It's not happening.

The present invention discloses the vaccine efficacy of Gn protein, Gc protein and NP protein.

An object of the present invention is to provide a vaccine composition against SFTS virus using Gn protein, Gc protein, and NP protein.

Other objects or specific objects of the present invention will be presented below.

As confirmed in the Examples below, the present inventors have prepared recombinant proteins (Gn-Fc and Gc-Fc) in which the extracellular domain of Gn or Gc protein is fused to the Fc region of a human antibody and NP protein (full-length sequence). protein) was individually immunized to type 1 interferon receptor KO mice, and whether protective immunity could be provided was compared and verified. As a result, all mice immunized with Gc-Fc protein died, but survival time increased statistically significantly compared to the control group (group immunized only with Fc), and 50% of mice immunized with Gn-Fc protein survived. , 60% of the mice immunized with the NP protein survived, confirming that each had a significant protective immunity providing effect. In addition, when Gn-Fc or a mixture of Gc-Fc and NP was immunized to type 1 interferon receptor KO mice to examine whether protective immunity was increased, 80% of the mixed immunization of Gc-Fc and NP survived, and Gn- In the case of mixed immunization of Fc and NP, 70% survived, and in the case of mixed immunization of Gn-Fc, Gc-Fc and NP, 60% survived, confirming that the overall survival rate of mixed immunization is higher than that of individual immunization. did

The present invention is provided based on these experimental results, and the vaccine composition of the present invention comprises, as an active ingredient, an extracellular domain of Gc protein, an extracellular domain of Gn protein, a full-length NP protein, or a mixture of two or more thereof, It relates to a vaccine composition against the SFTS virus.

In the present invention, in particular, the extracellular domain of the Gc protein or the extracellular domain of the Gn protein may be fused to a human antibody Fc region to improve in vivo half-life, and when fused to Fc, each fused protein interacts with each other. A hinge sequence with 2 to 15 amino acids can be inserted so as not to interfere with the formation of the three-dimensional structure. In addition, in order to produce these proteins using host cells by genetic recombination, a signal peptide sequence may be located at the N-terminus to facilitate separation and purification, as in the examples below. Since these sequences are removed when the protein is secreted to the outside of the cell, it does not exist in the isolated/purified protein.

As used herein, "vaccine" refers to prevention of infection or re-infection of the pathogen by inducing an immune response against the pathogen in a human host, reduction of severity of symptoms caused by the pathogen, or elimination of symptoms caused by the pathogen or the pathogen. It is meant to include substantial or complete elimination of the disease. Therefore, the vaccine composition of the present invention can be administered to humans prophylactically before infection with the corresponding pathogen or therapeutically administered to humans after infection with the corresponding pathogen. Here, the "immune response" is meant to include a humoral immune response, a cellular immune response, or both.

Also, in the present specification, "active ingredient" means a component capable of inducing an immune response alone or together with a carrier that cannot induce an immune response by itself. Therefore, the active ingredient itself does not necessarily have immunogenicity (the ability to induce an immune response). In the present invention, the active ingredient is preferably purified, although not essential. Here, “purified” refers to a state in which cell components of the original organism (ie, a transformed microorganism in the present invention) or its culture solution components (contaminants) in which the target substance exists are substantially reduced or removed. Substantially reduced or free of contaminants means a purity of 50% or more, preferably 90% or more, more preferably 99% or more. Purity can be evaluated through methods known in the art, such as chromatography, gel electrophoresis, and immunological analysis, and methods known in the art can be used as follows for purification methods.

The vaccine composition of the present invention can be prepared in any suitable, pharmaceutically acceptable formulation. For example, it may be in the form of an extemporaneous solution or suspension, a concentrated stock solution suitable for dilution prior to administration, or prepared in a reconstitutable form such as a lyophilized, freeze-dried, or frozen formulation.

The vaccine composition of the present invention may be formulated by including a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers that can be used for formulation of vaccine compositions are listed and regulated in the Korean Pharmacopoeia or other pharmacopeias, particularly in the US, Japan, and European pharmacopeias, and reference may be made to these pharmacopeias.

Such carriers will usually include diluents, excipients, stabilizers, preservatives, and the like. Suitable diluents include propylene glycol, polyethylene glycol, non-aqueous solvents such as vegetable oils such as olive oil and peanut oil, saline (preferably 0.8% saline), water containing a buffer medium (preferably 0.05M phosphate buffer), etc. suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, wheat flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, anhydrous skim milk , glycerol, propylene, glycol, water, ethanol, and the like, and suitable stabilizers include carbohydrates such as sorbitol, mannitol, starch, sucrose, dextran, glutamate, and glucose, or animal and vegetable substances such as milk powder, serum albumin, and casein. or proteins such as microbial proteins. Suitable preservatives include thimerosal, merthiolate, gentamicin, neomycin, nystatin, amphotericin B, tetracycline, penicillin, streptomycin, polymyxin B and the like.

The vaccine composition of the present invention may further include an antigen adjuvant. An antigenic adjuvant may consist of one or more substances that enhance an immune response to an antigen. Adjuvants can function as tissue depots that slowly release antigens and/or activators of the lymphatic system that non-specifically enhance immune responses (Hood et al., Immunology, Second Ed., 1984, benjamin/Cummings: Menlo Park , California, p.384). Antigenic adjuvants such as complete Freund's, incomplete Freund's, saponins, gelatinous aluminum adjuvants, surface active substances (eg lysolecithin, pluronic glycol, polyanions, peptides, oils or hydrocarbon emulsions, etc.), vegetable oils (cottonseed oil, peanut oil, corn oil, etc.), vitamin E acetate, and the like.

Currently, aluminum adjuvants are the most widely used among antigen adjuvants applicable to the human body, and these aluminum adjuvants are used in the gel form of aluminum salts such as aluminum phosphate, potassium aluminum sulfate, and aluminum hydroxide. there is. These aluminum adjuvants are generally known to elicit a Th2-type immune response to enhance vaccine efficacy (Sokolovska A et al., Vaccine. 2007 Jun 6;25(23):4575-85; O'Hagan DT AND Rappuoli R., Pharm Res. 2004 Sep;21(9):1519-30.) Invivogen's Alhydrogel® used in the examples of the present invention is a colloidal suspension of aluminum hydroxide. These aluminum adjuvants are well known in the art (R. Bomford. Immunological Adjuvants and Vaccines. NATO ASI Series 1989; 179: 35-41; Vogel FR AND Powell MF. Pharm Biotechnol. 1995; 6: 141- 228.; Derek T. O'Hagan, Methods in Molecular Medicine. 2000; April 15; 42: 65-90) It may be directly prepared and used, or a commercially distributed product may be purchased and used. Commercially distributed products include Invivogen's Alhydrogel® 2% product used in the examples below, Sigma's Aluminum hydroxide Gel product, BRENNTAG's Alhydrogel™ product, and the like.

The vaccine composition of the present invention can be prepared in arbitrary unit doses. A unit dose refers to the amount of an active ingredient and a pharmaceutically acceptable carrier contained in an individual packaged product for one or more administration to humans, and the appropriate amount of such active ingredient and carrier is An amount capable of functioning as a vaccine when administered one or more times can be determined nonclinically and clinically within the ordinary skill of a person skilled in the art.

The vaccine composition of the present invention can be preferably administered parenterally, such as rectally, transdermally, intravenously, intraarterially, intramuscularly, intradermally, subcutaneously, intraperitoneally, intraventricularly, etc. .

Vaccine compositions of the present invention can be administered as a controlled release system. Examples of such controlled-release systems include liposomes, implantable osmotic pumps, and transdermal patches. Preferably, it is the case of delivering an active ingredient in a liposomal manner. Regarding the delivery of active ingredients by liposome mode, see Langer, Science 249: 1527-1533 (1990), Treat at al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler. (des.), Liss, New York, pp. 353-365 (1989)] and the like. For other modes of delivery of active ingredients, see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987)], Buchwald et al., Surgery 88: 507 (1980), Saudek et al., N. Engl. J. Med. 321:574 (1989)], Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974), Controlled Drug Bioavailability. Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983)], During et al., Ann. Neurol. 25:351 (1989)], Howard et al., J. Neurosurg. 71:105 (1989)] and the like. These documents are considered part of this specification.

The dosage of the vaccine composition of the present invention may be determined by a medical person in consideration of patient characteristics such as age, weight, sex, symptoms, complications, and presence or absence of other diseases.

In addition, the time interval and frequency of administration may be determined in consideration of the dosage form used or the blood half-life of the active ingredient.

As such, the dose and the time interval or frequency of administration depend on the judgment of the medical practitioner and should be determined individually, but the usual appropriate dose will be determined within the range of about 0.1 to 10 mg per 1 kg of individual body weight per day, and the administration interval is 3 to 10 days, the number of administrations will be determined within the range of 1 to 5 times.

Each protein used as an active ingredient in the vaccine composition of the present invention can be prepared by DNA recombination technology known in the art using a gene encoding the protein. This method (i) prepares an expression vector containing the gene encoding each protein, (ii) transforms the expression vector into a host cell, (iii) cultures the transformed host cell, and finally (iv) ) isolating and purifying each of the above proteins from the culture. Genes encoding each of the above proteins are disclosed herein, and production of each corresponding protein by DNA recombination technology using these genes, specifically, control sequences such as promoters or terminators, signal peptide sequences, selection markers, etc. A considerable amount of literature has been accumulated in the art regarding the construction of expression vectors, recombinant DNA technology, transformation methods, culture methods after transformation, and the like, and these documents can be referred to for each relevant matter. Specifically, literature (Nucl. Acid Res. 14:5399-5467, 1986), literature (Tet. Lett. 27:5575-5578, 1986), literature (Nucl. Acid Res. 4:2557, 1977), literature (Lett. ., 28:2449, 1978), Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, (2001), F M Ausubel et al, Current Protocols in Molecular Biology, John Wiley amp Sons, Inc. (1994), Marston, F (1987) DNA Cloning Techniques, Cohen, S.N. et al., Proc. Natl. Acac. Sci. USA, 9:2110-2114 (1973). ), Hanahan, D., J. Mol. Biol., 166:557-580 (1983), Dower, W.J. et al., Nucleic. Acids Res., 16:6127-6145 (1988). (Capecchi, M.R., Cell, 22:479 (1980)), (Graham, F.L. et al., Virology, 52:456 (1973)), (Neumann, E. et al., EMBO J., 1: 841 (1982)), Wong, T.K. et al., Gene, 10:87 (1980), Gopal, Mol. Cell Biol., 5:1188-1190 (1985), Yang et al. ., Proc. Natl. Acad. Sci., 87:9568-9572 (1990) (Maniatis et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory (1982)) (Hitzeman et al., J Biol. Chem., 255, 12073-12080 (1980)), Luchansky et al Mol. Microbiol. 2, 637-646 (1988)), Sambrook et al., Molecular Cloning A Laboratory Manual, Cold Spring Harbor Laboratory Press, US (1989), Ausubel et al., Current Protocols in Molecular Biology, Jon Willey & Sons, US (1993)), by Sambrook, J. & Russel, D., Molecular Cloning, A LABORATORY MANUAL, Cold Spring Harbor Laboratory Press, Volumes 1 and 2, published January 15, 2001); can do.

As described above, the present invention can provide a vaccine composition against the SFTS virus, comprising the extracellular domain of the Gc protein, the extracellular domain of the Gn protein, the full-length NP protein, or a mixture thereof as an active ingredient.

1 is a diagram showing the structure and molecular weight of Gc protein, Gn protein or NP protein, which are active ingredients of the vaccine composition of the present invention.
2 is an SDS-PAGE result of Gc protein, Gn protein or NP protein, which are active ingredients of the vaccine composition of the present invention.
Figure 3 shows the results of specific antibody induction by Gn and Gc antigen immunization and virus neutralization ability test using immune serum (a. Antibody titer against Gn and Gn protein. b and c. Neutralization ability assay test against SFTS virus. FRNT : focus reduction neutralization titer)
4 shows the results of survival rate and body weight change after immunizing type 1 interferon receptor KO mice with Gc protein, Gn protein, or NP protein.
5 shows the results of survival rate and body weight change after immunizing type 1 interferon receptor KO mice with a mixture of Gc protein, Gn protein, and/or NP protein.

Hereinafter, the present invention will be described with reference to examples. However, the scope of the present invention is not limited to these examples.

<Example 1> Production of viral proteins to be used as vaccine antigens

<Example 1-1> Amino acid sequence of viral protein to be used as vaccine antigen

Using the amino acid information on GenBank, viral Gn protein, Gc protein and NP protein to be used as vaccine antigens were produced. Amino acid information on GenBank is GenBank ID: ATW63054.1 (Gn protein), GenBank ID: AOO85591.1 (Gc protein), GenBank ID: AJO16084.1 (NP protein).

For the Gn and Gc proteins, only signal peptide, transmembrane domain, and extracellular domain except for the cytoplasmic domain (SEQ ID NOs: 1 and 2, respectively) were used, and the full-length sequence (SEQ ID NO: 3) was used for the NP protein.

<Example 1-2> Virus protein gene cloning and expression

After synthesizing genes encoding Gn and Gc proteins (extracellular domain) (SEQ ID NOs: 4 and 5, respectively), the signal peptide gene (SEQ ID NO: 6) of human Ig kappa light chain is fused to the N-terminal as shown in FIG. 1, and human The recombinant gene in which the IgG1 CH2-CH3 Fc domain gene (SEQ ID NO: 7) was fused to the C-terminal via the hinge region (SEQ ID NO: 8) was cloned into pCEP4 mammalian expression vector. The gene (SEQ ID NO: 9) encoding the NP protein with 6 His (used for purification) linked to each of the N-terminal and C-terminal was cloned into a pET28a vector. The cloned gene was introduced into HEK293T cells (Gn, Gc) and E.coli BL21 (NP) to induce protein expression, and purified using Protein A/G column and Ni-NTA column, respectively. The size and purity of the purified protein was confirmed by SDS-PAGE (FIG. 2).

<Example 2> Immunity and antibody analysis of SFTS virus proteins

Gn-Fc (SEQ ID NO: 10, N-terminal DVAQAA amino acid sequence in SEQ ID NO: 10 is inserted for cloning), Gc-Fc (SEQ ID NO: 11, N-terminal DVAQAA amino acid sequence in SEQ ID NO: 10 is cloned) ), NP (SEQ ID NO: 12), and human Fc were immunized with type 1 interferon receptor KO mice (n = 4) three times at intervals of 2 weeks (20ug/mouse +Alum, Gn-Fc , Gc-Fc, Fc; 10ug/mouse + Alum, NP), blood was collected 2 weeks after the third immunization, and antibody titers against Gn, Gc, and NP were measured by ELISA (Fig. 3a). It was confirmed that high levels of antibodies were produced for each antigen (anti-Gn or anti-Gc IgG average titer = 1:4,096, anti-NP IgG average titer = 1:24,576; n = 4/group), especially NP antigen It was observed that antibody titers were highly induced. In order to confirm the virus neutralization ability of antibodies generated against viral glycoproteins (Gn and Gc), SFTS virus was reacted with immune serum, and focus reduction assay was performed by infecting Vero cells (Fig. 3 b and c) . Although almost similar levels of antibodies were produced, the serum obtained by immunizing Gn-Fc formed a slightly higher neutralizing antibody titer than the serum obtained by immunizing Gc-Fc, and both immunized sera obtained by immunizing only Fc It was confirmed that it had a significantly higher level of virus neutralizing ability (neutralizing antibody titer: Gn-Fc: 929 ± 1134, Gc-Fc: 209 ± 140, Fc: 42 ± 29; n = 4/group, mean titer ± S.D.).

<Example 3> Protective immune effect by SFTS virus protein immunization

Type 1 interferon receptor KO mice (n=4~6) were immunized three times at 2-week intervals with Gn-Fc, Gc-Fc, NP, and Fc (20ug/mouse +Alum, Gn-Fc, Gc-Fc , Fc; 10ug/mouse + Alum, NP), and 2 weeks after the third immunization, 10 ³ foci-forming units (FFU) of the SFTS virus were subcutaneously infected. And daily weight change and mortality were observed (FIG. 4).

Mice immunized with PBS (control group) gradually lost weight and all died by the 6th day of infection. Mice immunized with Fc (control group) gradually lost weight from the second day of infection and all died by the fifth day of infection. Mice immunized with Gc-Fc began to lose weight on the 4th day after infection, and all died on the 8th day after infection. Mice immunized with Gn-Fc began to lose weight on the 3rd day of infection, and half of them died by the 8th day of infection, but the rest of the mice recovered their body weight and survived. Mice immunized with NP began to lose weight from the third day of infection, and 40% died by the eighth day, and the remaining 60% mice gradually recovered their body weight and survived. When statistical analysis was performed (Log-rank (Mantel-Cox) Test), the effect of vaccine prolonging survival (Gc-Fc group: p = 0.0046) or reducing mortality (Gn-Fc group: p = 0.0054, NP Group: p = 0.0023) was confirmed to show a statistically significant improvement compared to the control group.

<Example 4> Protective immune effect by SFTS virus protein mixed immunization

A mixture of Gn-Fc and NP (Gn + NP, mixing ratio: 10ug + 10ug), a mixture of Gc-Fc and NP (Gc + NP, mixing ratio: 10ug + 10ug), a mixture of Gn-Fc and Gc-Fc and NP ( Gn+Gc+NP, mixing ratio: 10ug + 10ug + 10ug) and PBS (Control) were immunized with type 1 interferon receptor KO mice (n=7) three times at 2-week intervals (20 or 30ug/mouse, + Alum), and 2 weeks after the third immunization, 10 ³ foci-forming units (FFU) of SFTS virus were subcutaneously infected. And daily weight change and mortality were observed (FIG. 5).

Mice immunized with PBS (control group) gradually lost weight and all died by the 6th day of infection. Mice immunized with Gn+NP began to lose weight on the 4th day of infection, 14.3% died on the 5th day of infection and 28.6% died by the 6th day of infection, but the rest of the mice recovered their body weight and survived. Mice immunized with Gc+NP began to lose weight on the 5th day of infection, and 14.3% died on the 6th day of infection, but the rest of the mice recovered their body weight and survived. Mice immunized with Gn+Gc+NP began to lose weight on the 4th day, and 42.9% died on the 7th day of infection, but the rest of the mice recovered their weight and survived.

When statistical analysis was performed (Log-rank (Mantel-Cox) Test), the effect of vaccine prolonging survival (Gc-Fc group: p = 0.0046) or reducing mortality (Gn-Fc group: p = 0.0054, NP Group: p = 0.0023) was confirmed to show a statistically significant improvement compared to the control group.

<110> SNU R&DB Foundation <120> Vaccine Composition for SFTS virus <130> PP19-000-SNU-CHO <160> 12 <170> KoPatentIn 3.0 <210> 1 <211> 438 <212> PRT <213> Unknown < 220> <223> Extracellular domain of Gn protein of SFTS virus <400> 1 Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro Ile His Ser Asn Lys Ser 1 5 10 15 Ala Asp Ile Pro His Leu Leu Gly Tyr Ser Glu Lys Ile Cys Gln Ile 20 25 30 Asp Arg Leu Ile His Val Ser Ser Trp Leu Arg Asn His Ser Gln Phe 35 40 45 Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg Ser Gln Val Ser Tyr Tyr 50 55 60 Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser Gly Leu Leu Ser Pro Cys 65 70 75 80 Asp Ala Asp Trp Leu Gly Met Leu Val Val Lys Lys Ala Lys Gly Ser 85 90 95 Asp Met Ile Val Pro Gly Pro Ser Tyr Lys Gly Lys Val Phe Phe Glu 100 105 110 Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp Gly Cys Gly Ser Gly Lys 115 120 125 Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser Ser Asp Ser Gly Thr Ser 130 135 140 Ser Gly Leu Leu Pro Ser Asp Arg Val Leu Trp Ile Gly Asp Val Ala 145 150 155 160 Cys Gln Pro Met Thr Pro Ile Pro Glu Glu Thr Phe Leu Glu Leu Lys 165 170 175 Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile Cys Lys Ile Asp Gly Ile 180 185 190 Val Phe Asn Gln Cys Glu Gly Glu Ser Leu Pro Gln Pro Phe Asp Val 195 200 205 Ala Trp Met Asp Val Gly His Ser His Lys Ile Ile Met Arg Glu His 210 215 220 Lys Thr Lys Trp Val Gln Glu Ser Ser Ser Lys Asp Phe Val Cys Tyr 225 230 235 240 Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser Glu Lys Thr Cys Lys 245 250 255 Thr Ser Gly Ser Cys Arg Gly Asp Met Gln Phe Cys Lys Val Ala Gly 260 265 270 Cys Glu His Gly Glu Glu Ala Ser Glu Ala Lys Cys Arg Cys Ser Leu 275 280 285 Val His Lys Pro Gly Glu Val Val Val Ser Tyr Gly Gly Met Arg Val 290 295 300 Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met Met Ala Thr Leu Glu Val 305 310 315 320 Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys Thr Gly Cys His Leu Glu 325 330 335 Cys Ile Asn Gly Gly Val Arg Leu Ile Thr Leu Thr Ser Glu Leu Lys 340 345 350 Ser Ala Thr Val Cys Ala Ser His Phe Cys Ser Ser Ala Thr Ser Gly 355 360 365 Lys Lys Ser Thr Glu Ile Gln Phe His Ser Gly Ser Leu Val Gly Lys 370 375 380 Thr Ala Ile His Val Lys Gly Ala Leu Val Asp Gly Thr Glu Phe Thr 385 390 395 400 Phe Glu Gly Ser Cys Met Phe Pro Asp Gly Cys Asp Ala Val Asp Cys 405 410 415 Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro Gln Cys Tyr Pro Ala Lys 420 425 430 Lys Gly Gln Ala Gly Gln 435 <210> 2 <211> 473 <212> PRT <213> Unknown <220> <223> Extracellular domain of Gc protein of SFTS virus <400> 2 Cys Asp Glu Met Val His Ala Asp Ser Lys Leu Val Ser Cys Arg Gln 1 5 10 15 Gly Ser Gly Asn Met Lys Glu Cys Val Thr Thr Gly Arg Ala Leu Leu 20 25 30 Pro Ala Val Asn Pro Gly Gln Glu Ala Cys Leu His Phe Thr Ala Pro 35 40 45 Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile Lys Val Lys Arg Ile Asn 50 55 60 Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe Val Pro Asp Ala Arg Ser 65 70 75 80 Arg Cys Thr Ser Val Arg Arg Cys Arg Trp Ala Gly Asp Cys Gln Ser 85 90 95 Gly Cys Pro Pro His Phe Thr Ser Asn Ser Phe Ser Asp Asp Trp Ala 100 105 110 Gly Lys Met Asp Arg Ala Gly Leu Gly Phe Ser Gly Cys Ser Asp Gly 115 120 125 Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn Ala Ala Pro Ser Cys Ile 130 135 140 Phe Trp Arg Lys Trp Val Glu Asn Pro His Gly Ile Ile Trp Lys Val 145 150 155 160 Ser Pro Cys Ala Ala Trp Val Pro Ser Ala Val Ile Glu Leu Thr Met 165 170 175 Pro Ser Gly Glu Val Arg Thr Phe His Pro Met Ser Gly Ile Pro Thr 180 185 190 Gln Val Phe Lys Gly Val Ser Val Thr Tyr Leu Gly Ser Asp Met Glu 195 200 205 Val Ser Gly Leu Thr Asp Leu Cys Glu Ile Glu Glu Leu Lys Ser Lys 210 215 220 Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala Gly Met Gly Val Val Gly 225 230 235 240 Lys Val Gly Glu Ile Gln Cys Ser Ser Glu Glu Ser Ala Arg Thr Ile 245 250 255 Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp Leu Val Gly Ile Glu Leu 260 265 270 Arg Val Asp Asp Ala Val Cys Tyr Ser Lys Ile Thr Ser Val Glu Ala 275 280 285 Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr Ile Gly Gly Leu Arg Phe 290 295 300 Glu Arg Ser His Asp Ser Gln Gly Lys Ile Ser Gly Ser Pro Leu Asp 305 310 315 320 Ile Thr Ala Ile Arg Gly Ser Phe Ser Val Asn Tyr Arg Gly Leu Arg 325 330 335 Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys Thr Gly Glu Val Thr Asn 340 345 350 Val Ser Gly Cys Tyr Ser Cys Met Thr Gly Ala Lys Val Ser Ile Lys 355 360 365 Leu His Ser Ser Lys Asn Ser Thr Ala His Val Arg Cys Lys Gly Asp 370 375 380 Glu Thr Ala Phe Ser Val Leu Glu Gly Val His Ser Tyr Thr Val Ser 385 390 395 400 Leu Ser Phe Asp His Ala Val Val Asp Glu Gln Cys Gln Leu Asn Cys 405 410 415 Gly Gly His Glu Ser Gln Val Thr Leu Lys Gly Asn Leu Ile Phe Leu 420 425 430 Asp Val Pro Lys Phe Val Asp Gly Ser Tyr Met Gln Thr Tyr His Ser 435 440 445 Thr Val Pro Thr Gly Ala Asn Ile Pro Ser Pro Thr Asp Trp Leu Asn 450 455 460 Ala Leu Phe Gly Asn Gly Leu Ser Arg 465 470 <210> 3 < 211> 245 <212> PRT <213> Unknown <220> <223> NP protein of SFTS virus <400> 3 Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu 1 5 10 15 Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly 20 25 30 Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp 35 40 45 Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg 50 55 60 Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser 65 70 75 80 Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala 85 90 95 Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro 100 105 110 Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val 115 120 125 Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met 130 135 140 Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu 145 150 155 160 Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala 165 170 175 Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu 180 185 190 Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val 195 200 205 Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile 210 215 220 Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala 225 230 235 240 Ala Tyr Arg Asn Leu 245 <210> 4 <211> 1299 <212> DNA <213> Unknown <220> <223> gene encoding excellular protein domain of Gn of SFTS virus <400> 4 gactctggac ctattatctg tgctggacct attcattcaa acaaatctgc tgacatccct 60 catctgctgg gctactccga aaaaatctgc cagattgatc gactgatcca cgtgagctcc 120 tggctgcgga accatagcca gttccaggga tacgtcggac agagggg cgg gcgctctcag 180 gtgagttact atccagccga gaacagctac agccggtggt ccggactgct gtctccatgt 240 gacgcagatt ggct gggaat gctggtggtc aagaaagcta agggctcaga tatgattgtc 300 cccggcccta gctacaaggg gaaagtgttc tttgaacgcc cc accttcga cggctatgtg 360 ggctgggggt gcggatcagg caaaagccga acagagtccg gagaactgtg cagcagcgac 420 tctggaactt caagcggcct gctgcctagc gatagggtcc tgtggattgg cgacgtggcc 480 tgccagccaa tgaccccaat ccccgaggaa acatttctgg agctgaagag tttctcacag 540 a gcgaatttc cagacatctg caaaattgac ggcatcgtgt tcaaccagtg tgagggggaa 600 agcctgcctc agccatttga tgtcgcttgg atggacgtgg gccactccca taagatcatt 660 atgcgcgagc acaagactaa atgggtccag gaatcctcta gtaaggattt cgtgtgctac 7 20 aaagagggga ccggaccctg ttccgaatct gaggaaaaga cttgcaaaac cagtgggtca 780 tgtcggggag acatgcagtt ttgcaaggtg gctggctgg agcacgggga ggaagcctct 840 gaagctaaat gcagatgtag tctggtccat aaacccggcg aggtggtcgt gagttacgga 900 ggcatgcggg tgagacctaa atgctatggg ttcagcagaa tgatggcaac cctggaggtg 960 aac cagcctg aacagaggat cggacagtgc acaggctgtc acctggagtg tattaatggg 1020 ggagtccgac tgatcaccct gacaagtgaa ctgaagtcag caacagtgg cgcctctcat 1080 ttctgttcaa gcgccactag cggcaagaaa tccaccgaga ttcagtttca cagcgggtcc 1140 ctggtgggaa agactgctat ccatgtcaaa ggagcactgg tggatggcac agagttcact 1200 tttgaagggt cctgcatgtt tccagacgga tgtgatgcag tggactgcac attctgtcgc 1260 gagtttctga agaatcccca gtgctatcct gccaagaaa 1299 <210> 5 <211> 1419 <212> DNA <213> Unknown <220> <223> gene encoding excellular domain of Gc protein of SFTS virus <400> 5 tg cgatgaaa tggtccacgc cgatagcaaa ctggtcagct gtaggcaggg aagcgggaac 60 atgaaggaat gcgtcacaac cggaagagct ctgctgccag cagtgaaccc tggacaggag 120 gcctgcctgc acttcacagc tcctggaagt ccagactcaa aatgcctgaa gatcaaagtg 180 aagcggatta atctgaagtg taagaaaagc tcctcttact t tgtgcccga cgcaaggtca 240 cgctgcacta gcgtccggag atgtagatgg gcaggcgatt gccagagcgg atgtccacct 300 catttcacct ctaacagttt ttcagacgat tgggcaggca agatggacag ggcaggactg 360 gggttcagcg gatgctccga tggatgtgga ggagca gctt gcggatgttt caacgcagcc 420 ccctcctgca tcttttggcg caaatgggtg gagaatcctc acggcatcat ttggaaggtc 480 tccccttgtg ctgcatgggt gccatctgcc gtcatcgagc tgacaatgcc cagcggcgaa 540 gtgagaactt tccatccaat gtccgggatt cccacccagg tctttaaggg agtgtctgtc 600 acatatctgg gcagcgacat ggaggtgtcc gggctgactg atctgtgcga aatc gaggaa 660 ctgaaatcca agaaactggc cctggctcct tgtaatcagg caggaatggg agtggtcggc 720 aaggtcgggg agattcagtg cagttcagag gaaagcgcac gcaccatcaa gaaagacggg 780 tgtatttgga acgccgatct ggtgggaatc gagctgcgag tggacgatgc tgtctgctac 840 tccaaaatta catctgtgga agcagtcgcc aattattccg ctatccctac cacaattggc 900 gtggctgcta ttcatgtatg 1080 accggggcaa aagtgagtat taagctgcac agctccaaaa actcaacagc tcatgtgcgc 1140 tgtaagggag acgagactgc attctctgtg ctggaaggcg tccacagtta cacagtgtcc 1200 ctgtcttttg accatgccgt ggtcgatgag cagtgcc agc tgaactgtgg gggacacgaa 1260 agccaagtga ctctgaaagg caatctgatc ttcctggacg tgccaaagtt tgtcgatggg 1320 tcatacatgc agacctatca tagcactgtg cccaccggcg ccaatattcc atctcccaca 1380 gattggctga acgctctgtt tgggaatgga ctgagtcgg 1419 <210> 6 <211> 60 <212> DNA <213> Unknown <220> <223> gene encoding signal peptide <400> 6 atggagacag acacactcct g ctatgggta ctgctgctct gggttccagg ttccactggt 60 60 < 210> 7 <211> 654 <212> DNA <213> Unknown <220> <223> gene encoding human Fc <400> 7 gcacctgaac tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc caaggacacc 60 ctcatgatct cccggacccc tgaggtcaca tgcgtggtgg t ggacgtgag ccacgaagac 120 cctgaggtca agttcaactg gtacgtggac ggcgtggagg tgcataatgc caagacaaag 180 ccgcgggagg agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac cgtcctgcac 240 caggactggc tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc cctcccagcc 300 cccatcgaga aaaccatctc caaagccaaa gggcagcccc gagaaccaca ggtgtacacc 360 ctgcccccat cccgggatga gctgaccaag aaccaggtca gcctgacctg cctggtcaaa 420 ggcttctatc ccagcgacat cgccgtggag tgggagca atgggcagcc ggagaacaac 480 tacaagacca cgcctcccgt gctggactcc gacggctcct tcttcctcta cagcaagctc 540 accgtggaca agagcaggtg gcagcagggg aacgtcttct catgctccgt gatgcatgag 600 gctctgcaca accactacac gcagaagagc ctctccctgt ccccgggtaa atga 654 <210> 8 <211> 60 <212> DNA <213> Unknown <220> <223> gene encoding hinge region <400> 8 ggccaggccg gccaggagcc caaatctagc gacaaaactc acacaagccc accgagccca 60 60 <2 10> 9 < 211 > 738 <212> DNA <213> Unknown <220> <223> gene encoding NP protein of SFTS virus <400> 9 atgtcggagt ggtccaggat tgcagtggag tttggtgagc agcagctcaa tttgtctgag 60 cttgaggatt tcgcgagaga actggcctat gaaggccttg accctg cttt gatcatcaag 120 aagctgaagg agacaggtgg agatgattgg gtgaaggata caaagttcat cattgtcttt 180 gccctgactc gaggcaacaa gatcgtcaag gcatcaggga aaatgtcaaa ctcagggtcc 240 aagaggttga tggcactcca agagaaatat ggactggttg agagggcaga gaccaggctc 300 tcaatcactc ctgtgagggt tgcgcagagc cttcccactt ggacatgtgc tgcagcagca 360 gccttga agg agtatctccc tgtggggcca gccgtcatga acctgaaggt cgagaattat 420 ccccctgaga tgatgtgcat ggcctttgga tccctgattc caactgcggg ggtatctgaa 480 gccacaacca agaccctgat ggaggcctac tctctgtggc aagatgcctt caccaagact 540 atcaat gtga agatgcgcgg agccagcaag acagaagttt acaactcctt cagggaccct 600 cttcatgctg ctgtgaactc tgtcttcttt cccaatgatg ttcgggtaaa gtggctgaag 660 gccaagggga tccttggccc agatggggtc cccagcagag ctgctgaggt tgctgctgct 720 gcttacagaa acctgtaa 738 <210> 10 <211> 676 <212> PRT <213> Unknown <220 > <223> Gn-Fc <400> 10 Asp Val Ala Gln Ala Ala Asp Ser Gly Pro Ile Ile Cys Ala Gly Pro 1 5 10 15 Ile His Ser Asn Lys Ser Ala Asp Ile Pro His Leu Leu Gly Tyr Ser 20 25 30 Glu Lys Ile Cys Gln Ile Asp Arg Leu Ile His Val Ser Ser Trp Leu 35 40 45 Arg Asn His Ser Gln Phe Gln Gly Tyr Val Gly Gln Arg Gly Gly Arg 50 55 60 Ser Gln Val Ser Tyr Tyr Pro Ala Glu Asn Ser Tyr Ser Arg Trp Ser 65 70 75 80 Gly Leu Leu Ser Pro Cys Asp Ala Asp Trp Leu Gly Met Leu Val Val 85 90 95 Lys Lys Ala Lys Gly Ser Asp Met Ile Val Pro Gly Pro Ser Tyr Lys 100 105 110 Gly Lys Val Phe Phe Glu Arg Pro Thr Phe Asp Gly Tyr Val Gly Trp 115 120 125 Gly Cys Gly Ser Gly Lys Ser Arg Thr Glu Ser Gly Glu Leu Cys Ser 130 135 140 Ser Asp Ser Gly Thr Ser Ser Gly Leu Leu Pro Ser Asp Arg Val Leu 145 150 155 160 Trp Ile Gly Asp Val Ala Cys Gln Pro Met Thr Pro Ile Pro Glu Glu 165 170 175 Thr Phe Leu Glu Leu Lys Ser Phe Ser Gln Ser Glu Phe Pro Asp Ile 180 185 190 Cys Lys Ile Asp Gly Ile Val Phe Asn Gln Cys Glu Gly Glu Ser Leu 195 200 205 Pro Gln Pro Phe Asp Val Ala Trp Met Asp Val Gly His Ser His Lys 210 215 220 Ile Ile Met Arg Glu His Lys Thr Lys Trp Val Gln Glu Ser Ser Ser Ser 225 230 235 240 Lys Asp Phe Val Cys Tyr Lys Glu Gly Thr Gly Pro Cys Ser Glu Ser 245 250 255 Glu Glu Lys Thr Cys Lys Thr Ser Gly Ser Cys Arg Gly Asp Met Gln 260 265 270 Phe Cys Lys Val Ala Gly Cys Glu His Gly Glu Glu Ala Ser Glu Ala 275 280 285 Lys Cys Arg Cys Ser Leu Val His Lys Pro Gly Glu Val Val Val Ser 290 295 300 Tyr Gly Gly Met Arg Val Arg Pro Lys Cys Tyr Gly Phe Ser Arg Met 305 310 315 320 Met Ala Thr Leu Glu Val Asn Gln Pro Glu Gln Arg Ile Gly Gln Cys 325 330 335 Thr Gly Cys His Leu Glu Cys Ile Asn Gly Gly Val Arg Leu Ile Thr 340 345 350 Leu Thr Ser Glu Leu Lys Ser Ala Thr Val Cys Ala Ser His Phe Cys 355 360 365 Ser Ser Ala Thr Ser Gly Lys Lys Ser Thr Glu Ile Gln Phe His Ser 370 375 380 Gly Ser Leu Val Gly Lys Thr Ala Ile His Val Lys Gly Ala Leu Val 385 390 395 400 Asp Gly Thr Glu Phe Thr Phe Glu Gly Ser Cys Met Phe Pro Asp Gly 405 410 415 Cys Asp Ala Val Asp Cys Thr Phe Cys Arg Glu Phe Leu Lys Asn Pro 420 425 430 Gln Cys Tyr Pro Ala Lys Lys Gly Gln Ala Gly Gln Glu Pro Lys Ser 435 440 445 Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro Glu Leu Leu 450 455 460 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 465 470 475 480 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 485 490 495 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 500 505 510 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 515 520 525 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 530 535 540 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 545 550 555 560 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 565 570 575 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 580 585 590 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 595 600 605 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 610 615 620 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 625 630 635 640 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 645 650 655 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 660 665 670 Ser Pro Gly Lys 675 <210> 11 <211> 711 <212> PRT <213> Unknown <220> <223> Gc-Fc <400> 11 Asp Val Ala Gln Ala Ala Cys Asp Glu Met Val His Ala Asp Ser Lys 1 5 10 15 Leu Val Ser Cys Arg Gln Gly Ser Gly Asn Met Lys Glu Cys Val Thr 20 25 30 Thr Gly Arg Ala Leu Leu Pro Ala Val Asn Pro Gly Gln Glu Ala Cys 35 40 45 Leu His Phe Thr Ala Pro Gly Ser Pro Asp Ser Lys Cys Leu Lys Ile 50 55 60 Lys Val Lys Arg Ile Asn Leu Lys Cys Lys Lys Ser Ser Ser Tyr Phe 65 70 75 80 Val Pro Asp Ala Arg Ser Arg Cys Thr Ser Val Arg Arg Cys Arg Trp 85 90 95 Ala Gly Asp Cys Gln Ser Gly Cys Pro Pro His Phe Thr Ser Asn Ser 100 105 110 Phe Ser Asp Asp Trp Ala Gly Lys Met Asp Arg Ala Gly Leu Gly Phe 115 120 125 Ser Gly Cys Ser Asp Gly Cys Gly Gly Ala Ala Cys Gly Cys Phe Asn 130 135 140 Ala Ala Pro Ser Cys Ile Phe Trp Arg Lys Trp Val Glu Asn Pro His 145 150 155 160 Gly Ile Ile Trp Lys Val Ser Pro Cys Ala Ala Trp Val Pro Ser Ala 165 170 175 Val Ile Glu Leu Thr Met Pro Ser Gly Glu Val Arg Thr Phe His Pro 180 185 190 Met Ser Gly Ile Pro Thr Gln Val Phe Lys Gly Val Ser Val Thr Tyr 195 200 205 Leu Gly Ser Asp Met Glu Val Ser Gly Leu Thr Asp Leu Cys Glu Ile 210 215 220 Glu Glu Leu Lys Ser Lys Lys Leu Ala Leu Ala Pro Cys Asn Gln Ala 225 230 235 240 Gly Met Gly Val Val Gly Lys Val Gly Glu Ile Gln Cys Ser Ser Glu 245 250 255 Glu Ser Ala Arg Thr Ile Lys Lys Asp Gly Cys Ile Trp Asn Ala Asp 260 265 270 Leu Val Gly Ile Glu Leu Arg Val Asp Asp Ala Val Cys Tyr Ser Lys 275 280 285 Ile Thr Ser Val Glu Ala Val Ala Asn Tyr Ser Ala Ile Pro Thr Thr 290 295 300 Ile Gly Gly Leu Arg Phe Glu Arg Ser His Asp Ser Gln Gly Lys Ile 305 310 315 320 Ser Gly Ser Pro Leu Asp Ile Thr Ala Ile Arg Gly Ser Phe Ser Val 325 330 335 Asn Tyr Arg Gly Leu Arg Leu Ser Leu Ser Glu Ile Thr Ala Thr Cys 340 345 350 Thr Gly Glu Val Thr Asn Val Ser Gly Cys Tyr Ser Cys Met Thr Gly 355 360 365 Ala Lys Val Ser Ile Lys Leu His Ser Ser Lys Asn Ser Thr Ala His 370 375 380 Val Arg Cys Lys Gly Asp Glu Thr Ala Phe Ser Val Leu Glu Gly Val 385 390 395 400 His Ser Tyr Thr Val Ser Leu Ser Phe Asp His Ala Val Val Asp Glu 405 410 415 Gln Cys Gln Leu Asn Cys Gly Gly His Glu Ser Gln Val Thr Leu Lys 420 425 430 Gly Asn Leu Ile Phe Leu Asp Val Pro Lys Phe Val Asp Gly Ser Tyr 435 440 445 Met Gln Thr Tyr His Ser Thr Val Pro Thr Gly Ala Asn Ile Pro Ser 450 455 460 Pro Thr Asp Trp Leu Asn Ala Leu Phe Gly Asn Gly Leu Ser Arg Glu 465 470 475 480 Pro Lys Ser Ser Asp Lys Thr His Thr Ser Pro Pro Ser Pro Ala Pro 485 490 495 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 500 505 510 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 515 520 525 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 530 535 540 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 545 550 555 560 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 565 570 575 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 580 585 590 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 595 600 605 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 610 615 620 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 625 630 635 640 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 645 650 655 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 660 665 670 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 675 680 685 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 690 695 700 Leu Ser Leu Ser Pro Gly Lys 705 710 <210> 12 <211> 245 <212> PRT <213> Unknown <220> <223> NP < 400> 12 Met Ser Glu Trp Ser Arg Ile Ala Val Glu Phe Gly Glu Gln Gln Leu 1 5 10 15 Asn Leu Ser Glu Leu Glu Asp Phe Ala Arg Glu Leu Ala Tyr Glu Gly 20 25 30 Leu Asp Pro Ala Leu Ile Ile Lys Lys Leu Lys Glu Thr Gly Gly Asp 35 40 45 Asp Trp Val Lys Asp Thr Lys Phe Ile Ile Val Phe Ala Leu Thr Arg 50 55 60 Gly Asn Lys Ile Val Lys Ala Ser Gly Lys Met Ser Asn Ser Gly Ser 65 70 75 80 Lys Arg Leu Met Ala Leu Gln Glu Lys Tyr Gly Leu Val Glu Arg Ala 85 90 95 Glu Thr Arg Leu Ser Ile Thr Pro Val Arg Val Ala Gln Ser Leu Pro 100 105 110 Thr Trp Thr Cys Ala Ala Ala Ala Ala Leu Lys Glu Tyr Leu Pro Val 115 120 125 Gly Pro Ala Val Met Asn Leu Lys Val Glu Asn Tyr Pro Pro Glu Met 130 135 140 Met Cys Met Ala Phe Gly Ser Leu Ile Pro Thr Ala Gly Val Ser Glu 145 150 155 160 Ala Thr Thr Lys Thr Leu Met Glu Ala Tyr Ser Leu Trp Gln Asp Ala 165 170 175 Phe Thr Lys Thr Ile Asn Val Lys Met Arg Gly Ala Ser Lys Thr Glu 180 185 190 Val Tyr Asn Ser Phe Arg Asp Pro Leu His Ala Ala Val Asn Ser Val 195 200 205 Phe Phe Pro Asn Asp Val Arg Val Lys Trp Leu Lys Ala Lys Gly Ile 210 215 220 Leu Gly Pro Asp Gly Val Pro Ser Arg Ala Ala Glu Val Ala Ala Ala 225 230 235 240 Ala Tyr Arg Asn Leu 245

Claims (8)

A mixture of the extracellular domain of the Gn protein (SEQ ID NO: 1) and the full-length NP protein (SEQ ID NO: 3) as an active ingredient,
The Gn protein is in the form of a fusion to the Fc region of a human antibody,
Vaccine composition against SFTS virus.
In paragraph 1,
The extracellular domain of the Gn protein and the Fc region of the human antibody are fused with a hinge sequence of 2 to 15 amino acids interposed between these proteins so as not to interfere with the formation of a three-dimensional structure between these proteins. Vaccine composition characterized by.
In paragraph 1,
The vaccine composition in which the Gn protein is fused to the Fc region of a human antibody is composed of a sequence excluding the N-terminal DVAQAA (Asp Val Ala Gln Ala Ala) amino acid sequence from the amino acid sequence of SEQ ID NO: 10 .
delete According to claim 1,
The vaccine composition, characterized in that the composition comprises a pharmaceutically acceptable carrier.
According to claim 5,
The pharmaceutically acceptable carrier is a vaccine composition, characterized in that it comprises at least one selected from the group consisting of diluents, excipients, stabilizers and preservatives.
According to claim 1,
The composition is a vaccine composition, characterized in that it further comprises an active ingredient adjuvant.
According to claim 7,
The adjuvant is a vaccine composition, characterized in that the aluminum salt in the form of a gel.

Images